• Muhammad Nauman Sheikh
  • Sajid Hanif
  • Majid Zia
  • Zahur Qayyum


Background: The extensive use of tobacco and its associated problematic health issues have been aconcern to mankind. The World Health Organization (WHO) estimates that approximately one-third ofthe global population aged 15 years or older are smokers and each smoker consumes an average of 15cigarettes daily. The objective of this study was to establish the effect of nicotine on an in vitroreconstituted oral mucosa model, the effect of treatment with this compound was measured in terms ofcytokine production. Method: Observational laboratory based study design was used to carry out theexperiment. The reconstituted human epithelium model used in the study was prepared and supplied bySkin Ethic Laboratories, Nice, France. The effect of nicotine on epithelial cytokine production wasassessed using commercially available assay kits (R&D systems). This was done using the enzymelinked immuno-sorbent assay. Result: In this study there was evidence that after 5 minutes treatment onun-inflamed mucosa with nicotine at 10 mm concentration GM-CSF release decreased, and also after 24hours treatment with nicotine at 10mM concentration GM-CSF release increased. TNF-α increasedrelease of pro-inflammatory cytokines, IL-6, IL-8, and also GM-CSF from the model mucosa after 24hours, but had no effect on the release of IL-1α., IL-6, IL-8, and GM-CSF after 5 minutes and 24 hoursrespectively. Conclusion: In conclusion at all the concentrations used in this experiment, nicotine had noeffect on the TNF-α stimulated tissue and un-inflamed mucosa and had no significant effect on cytokinerelease including IL-1α., IL-6, IL-8, and GM-CSF after 5 minutes and 24 hours respectively.Keywords: Nicotine, oral mucosa, cytokine


WHO, Tobacco or health: a global status report. Geneva: World

Health Organization 1991 p. 1–48.

Mohamed S Al Moamary. Tobacco consummation: Is it still a

dilemma? Ann Thorac Med 2010;5(4):193–4.

Mannino DM. Chronic obstructive pulmonary disease: definition

and epidemiology. Respir Care 2003;48:1185–91, discussion


Tsiara S, Elisaf M, Mikhailidis DP. Influence of smoking on

predictors of vascular disease. Angiology 2003;54:507–30.

Tanski SE, Prokhorov AV, Klein JD. Youth and tobacco. Minerva

Pediatr 2004;56:553–6.

Lu GH, Ralapati S. Application of high-performance capillary

electrophoresis to the quantitative analysis of nicotine and

profiling of other alkaloids in ATF-regulated tobacco products.

Electrophoresis 1998;19(1):19–26.

Baldwin IT. Mechanism of damage-induced alkaloid production

in wild tobacco. J Chem Ecology 1989;15:1661–80.

Hashimoto T, Yamada Y. Alkaloid biogenesis: molecular aspects.

Annual Review of Plant Physiology and Plant Molecular Biology


Taybos G. Oral changes associated with tobacco use. Am J Med

Sci 2003;326:179–82.

Sham AS, Cheung LK, Jin LJ, Corbet EF. The effects of tobacco

use on oral health. Hong Kong Med J 2003;9:271–7.

Mirbod SM, Ahing SI. Tobacco-associated lesions of the oral

cavity: Part I. Nonmalignant lesions. J Can Dent Assoc


Dinarello CA. Interleukin-1, interleukin-1 receptors and interleukin-

receptor antagonist. Int Rev Immunol 1998;16:457–99.

Kupper TS, Min K, Sehgal P, Mizutani H, Birchall N, Ray A,

May L. Production of IL-6 by keratinocytes. Implications for

epidermal inflammation and immunity. Ann N Y Acad Sci

;557:454–64, discussion 464–5.

Taub DD, Anver M, Oppenheim JJ, Longo DL, Murphy WJ. T

lymphocyte recruitment by interleukin-8 (IL-8). IL-8-induced

degranulation of neutrophils releases potent chemoattractants for

human T lymphocytes both in vitro and in vivo. J Clin Invest


Li J, Ireland GW, Farthing PM, Thornhill MH. Epidermal and oral

keratinocytes are induced to produce RANTES and IL-8 by

cytokine stimulation. J Invest Dermatol 1996;106:661–6.

Braunstein S, Kaplan G, Gottlieb AB, Schwartz M, Walsh G,

Abalos RM, et al. GM-CSF activates regenerative epidermal

growth and stimulates keratinocyte proliferation in human skin in

vivo. J Invest Dermatol 1994;103:601–4.

Lee JH, Kim NS, Kwon TH, Jang YS, Yang MS. Increased

production of human granulocyte-macrophage colony stimulating

factor (hGM-CSF) by the addition of stabilizing polymer in plant

suspension cultures. J Biotechnol 2002;96(3):205–11.

Klapproth H, Racké K, Wessler I. Acetylcholine and nicotine

stimulate the release of granulocyte-macrophage colony

stimulating factor from cultured human bronchial epithelial cells.

Naunyn Schmiedebergs Arch Pharmacol 1998;357:472–5.

Rioux N, Castonguay A. 4-(methylnitrosamino)-1-(3-pyridyl)-1-

butanone modulation of cytokine release in U937 human

macrophages. Cancer Immunol Immunother 2001;49:663–70.

Kwon OS, Chung JH, Cho KH. Nicotine enhanced epithelial

differentiation in reconstructed human oral mucosa in vitro. Skin

Pharmacol Appl Skin Physio 1999;12:227–34.

Hoffmann D, Hoffmann I. The changing cigarette, 1950-1995. J

Toxicol Environ Health 1997;50(4):307–64.

Hecht SS. Biochemistry, biology, and carcinogenicity of tobaccospecific N-nitrosamines. Chem Res Toxicol 1998;11:559–603.

Squier CA, Johnson GK. Role of nicotine as a cofactor in

smokeless tobacco carcinogenesis. Smoking and Tobacco Control

Monograph 2. NIH Pub 92;1992.p. 153–60.

Dussor GO, Leong AS, Gracia NB, Kilo S, Price TJ, Hargreaves

KM. Potentiation of evoked calcitonin gene- related peptide release

from oral mucosa: a potential basis for the pro- inflammatory

effects of nicotine. Eur J Neuro Sci 2003;18:2515–26.

Barker JN, Mitra RS, Griffiths CE, Dixit VM, Nickoloff BJ.

Keratinocytes as initiators of inflammation. Lancet


Johnson GK, Organ CC. Prostaglandin E2 and interleukin-1

concentrations in nicotine-exposed oral keratinocyte cultures. J

Periodontal Res 1997;32:447–54.

Rupniak HT, Rowlatt C, Lane EB, Steele JG, Trejdosiewicz LK,

Laskiewicz B, Povey S, Hill BT. Characteristics of four new

human cell lines derived from squamous cell carcinomas of the

head and neck. J Natl Cancer Inst 1985;75(4):621–35.

Rosdy M, Clauss LC. Terminal epidermal differentiation of

human keratinocytes grown in chemically defined medium on

inert filter substrates at the air-liquid interface. J Invest Dermatol


Christen AG. The case against smokeless tobacco: five facts for

the health professional to consider. J Am Dent Assoc


Yamamoto T, Osaki T, Yoneda K, Ueta E. Cytokine production

by keratinocytes and mononuclear infiltrates in oral lichen planus.

J Oral Pathol Med 1994;23:309–15.

Kupper TS, Lee F, Birchall N, Clark S, Dower S. Interleukin 1

binds to specific receptors on human keratinocytes and induces

granulocyte macrophage colony-stimulating factor mRNA and

protein. A potential autocrine role for interleukin 1 in epidermis. J

Clin Invest 1988;82:1787–92.

Partridge M, Chantry D, Turner M, Feldmann M. Production of

interleukin-1 and interleukin-6 by human keratinocytes and

squamous cell carcinoma cell lines. J Invest Dermatol


Cheng YA, Tsai CC. Nicotine- and arecoline-induced interleukin-

secretion and intercellular adhesion molecular-1 expression in

human oral epidermoid carcinoma cells in vitro. Arch Oral Biol


Nakano Y, Kobayashi W, Sugai S, Kimura H, Yagihashi S.

Expression of tumor necrosis factor-alpha and interleukin-6 in oral

squamous cell carcinoma. Jpn J Cancer Res 1999;90:858–66.

Hong SH, Ondrey FG, Avis IM, Chen Z, Loukinova E, Cavanaugh

PF Jr, et al. Cyclooxygenase regulates human oropharyngeal

carcinomas via the proinflammatory cytokine IL-6: a general role

for inflammation? FASEB J 2000;14(11):1499–507.

Wendell KJ, Stein SH. Regulation of cytokine production in

human gingival fibroblasts following treatment with nicotine and

lipopolysaccharide. J Periodontol 2001;72:1038–44.

Sugiyama A, Uehara A, Iki K, Matsushita K, Nakamura R,

Ogawa T, et al. Activation of human gingival epithelial cells by

cell-surface components of black-pigmented bacteria:

augmentation of production of interleukin-8, granulocyte colonystimulating factor and granulocyte-macrophage colonystimulating factor and expression of intercellular adhesion

molecule 1. J Med Microbiol 2002;51:27–33.

Gabri MR, Menna PL, Scursoni AM, Gomez DE, Alonso DF.

Role of tumor-derived granulocyte-macrophage colonystimulating factor in mice bearing a highly invasive and metastatic

mammary carcinoma. Pathobiology 1999;67:180–5.

Jablonska E, Piecuch J, Piotrowski L, Grabowska Z. GM-CSF in

the culture supernatants of neutrophils and serum levels in patients

with inflammation and squamous cell carcinoma of oral cavity.

Pol Merkuriusz Lek. 2001;11:394–7.